Volume 18 Issue 4
Aug.  2022
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MA Yanping, HAO Le, FENG Guoqing, LIANG Zhiling, MA Jiangyao, KE Hao, LIU Zhenxing. Construction and characterization research of essA, essB and essC-deleted mutants in Streptococcus agalactiae from tilapia[J]. South China Fisheries Science, 2022, 18(4): 76-86. DOI: 10.12131/20210246
Citation: MA Yanping, HAO Le, FENG Guoqing, LIANG Zhiling, MA Jiangyao, KE Hao, LIU Zhenxing. Construction and characterization research of essA, essB and essC-deleted mutants in Streptococcus agalactiae from tilapia[J]. South China Fisheries Science, 2022, 18(4): 76-86. DOI: 10.12131/20210246
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Construction and characterization research of essA, essB and essC-deleted mutants in Streptococcus agalactiae from tilapia

  • Institute of Animal Health, Guangdong Academy of Agricultural Sciences/Key Laboratory of Livestock Disease Prevention of Guangdong Province/Scientific Observation and Experiment Station of Veterinary Drugs and Diagnostic Techniques of Guangdong Province, Ministry of Agriculture and Rural Affairs, Guangzhou 510640, China

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  • Received Date: August 25, 2021
  • Revised Date: December 13, 2021
  • Accepted Date: December 20, 2021
  • Available Online: February 05, 2022
    Graphical Abstract
    • Abstract
  • The membrane proteins are critical to substrate protein secretion in VII secretion system existing in Streptococcus agalactiae. In this study, we constructed the membrane proteins knockout vectors with chloramphenicol tag including pSET4s-ΔessA, pSET4s-ΔessB and pSET4s-ΔessC, and after the homologous recombination, we screened the mutant S. agalactiae ΔessA, ΔessB and ΔessC via PCR assays. Based on the growth curve analysis, S. agalactiae ΔessA, ΔessB and ΔessC strains showed significantly slower growth process than the wild type strain (WT). ΔessA and ΔessB strains had significant difference in early stage of growth compared with WT strain (P<0.01). According to the secretion product analysis of ESAT6 protein, S. agalactiae ΔessA, ΔessB and ΔessC strains showed significantly lower ESAT6 product than WT (P<0.01). According to challenge test analysis, the deletion of essA, essB or essC gene significantly reduced virulence of S. agalactiae ΔessA, ΔessB and ΔessC compared with WT (P<0.01). The results suggest that essA, essB and essC proteins are important component membrane proteins of VII secretion system in S. agalactiae, and these genes deletions cause ESAT6 mRNA expression and virulence decline.
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    • References (31)
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